Objective To study the chemical constituents of the EtOAc soluble fraction obtained from the stems and branches of Euonymus alatus with anti-myocardial ischemic effect. Methods Chemical constituents were isolated and purified by solvent extraction together with various chromatographic techniques, and the structures were identified by physicochemical properties and spectral analyses. ResultsFive compounds, 1-[3-?-D-glucopyranosyloxy)-4, 5-dihydroxyphenyl]-ethanone (Ⅰ), phenethyl alcohol 8-O-?-D-glucopyranosyl-(1→2)-?-D-glucopyranoside (Ⅱ), eugenyl-O-?-D-glucopyranoside (Ⅲ), hyperin (Ⅳ), and hesperidin (Ⅴ) were isolated. Conclusion Compound Ⅰ is determined as a new compound named guijianyuside. Compounds Ⅱ and Ⅲ are isolated from this plant for the first time.

With the progess in studying gene structure and function of foot and mouth disease virus (FMDV), FMDV can express exogenous genes in different sites. Through transforming and modifying FMDV can achieve different application purposes such as improving virus titer, introducing tag, improving immune responses, and reducing pathogenicity. From the perspective of FMDV receiving inserted exogenous gene, this paper mainly describes the latest relevant developments of FMDV's expression to exogenous gene.
Foot-and-Mouth Disease Virus ; genetics ; Genetic Engineering ; Mutagenesis, Insertional

This study was aimed to optimize the purification technology for Sambucus williamsii Hance. With the morroniside as a marker, the purification technology for Sambucus williamsii Hance was optimized by different types of macroporous resin. The results showed that the optimum purification technology was that, the extract of less than 1:250 (morroniside:resin) was adsorbed and the AB-8 resin was washed with distilled water, and then the morroni-side was eluted from the macroporous resin with 10% ethanol. And the content of the morroniside was more than 50%. It was concluded that the purification technology was simple, reliable, repeatable and suitable for industrial production.

To generate an epitope-mutated foot-and-mouth disease virus (FMDV) as a marker vaccine, the infectious clone pAsia 1-FMDV containing the complete genomic cDNA of Asia 1 type FMDV was used as backbone, the residues at positions 27 and 31 in the 3D gene were mutated (H27Y and N31R). The resulting plasmid pAsia 1-FMDV-3DM encoding a mutated epitope was transfected into BHK-21 cells and the recombinant virus rAsia 1-3DM was rescued. The recombinant virus showed similar biological characteristics comparable with the parental virus. In serological neutralization test the antisera against recombine virus have a good reactivity with parental virus. The antisera against the mutant virus were shown to be reactive with the mutated epitope but not the wild-type one. The results indicated that the two virus strains could be distinguished by western blotting using synthetic peptides. This epitope-mutated FMDV strain will be evaluated as a potential marker vaccine against FMDV infections.
Animals ; Cell Line ; DNA, Complementary ; Epitopes ; genetics ; Foot-and-Mouth Disease Virus ; genetics ; Immune Sera ; Neutralization Tests ; Transfection

Objective To observe the protective effects of various extracts from Flos Daturae (FD), including active fraction (AF-FD), withanolides constituents (WC-FD), and flavonoids constituents (FC-FD) on Chinese hamster ovary (CHO) cells against cytotoxicity induced by DMSO. Methods The survival rate of CHO cells was examined by MTT assay and LDH leakage assays. Results Using MTT assay, coincubation of CHO cells with 3% DMSO for 24 h resulted in a significant reduction of survival rate of CHO cells. AF-FD was tested in a range of 10—80 ?g/mL to improve the survival rate of CHO cells in a dose-dependent manner. FC-FD (2.5—20 ?g/mL), but not WC-FD (30—120 ?g/mL), could significantly relieve the injury induced by 3% DMSO in CHO cells. In the measurement of LDH leakage, coincubation of CHO cells with 4.5% DMSO for 24 h obviously increased LDH release. However, all the compounds tested, including AF-FD (10—80 ?g/mL), WC-FD (30—120 ?g/mL), and FC-FD (2.5—20 ?g/mL) had no effect on LDH leakage induced by 4.5% DMSO. Conclusion The findings suggest that the FC-FD may protect CHO cells from DMSO cytotoxicity assessed by MTT assay, which may be associated with improving mitochondrial function, but not protecting the membrane injury of CHO cells.

Objective To identify Isatidis Radix in different areas by SDS-PAGE;To provide the basis for the identification and quality evaluation of Isatidis Radix. Methods By using SDS-PAGE technology, the protein profiles of Isatidis Radix in Heilongjiang and Hebei regions were established. Results Isatidis Radix protein in Heilongjiang contained about 13 protein subunits, and the major subunits were 10.5 kD, 23.0 kD, 24.9 kD, and 44.1 kD. Isatidis Radix protein in Hebei contained about 12 subunits, and the major subunits were 55.6 kD, 45.9 kD, 34.3 kD, 18.9 kD, 12.4 kD, and 10.5 kD. Conclusion SDS-PAGE technology can be used as one of the references to identify Isatidis Radix in different areas.

AIM To study the flavonoids from the leaves of Astragalus membranaceus (Fish.) Bge..METHODS The ethyl acetate and n-butyl alcohol fractions of 75％ ethanol extract from A.membranaceus leaves were isolated and purified by silica,ODS and preparative HPLC column,then the structures of obtained compounds were identified by spectral data.RESULTS Thirteen compounds were isolated and identified as quercetin (1),quercetin-3-O-β-D-glucopyranoside (2),rhamnocitrin-3-O-β-D-glucopyranoside (3),rhamnocitrin-3-O-β-neohesperidoside (4),rhamnocitrin-3-O-3-D-glucopyranoside (1'''→2'')-β-D-apiofuranosyl (5),complanatuside (6),glycitein (7),4',7-dihydroxy-3-methoxy isoflavone (8),genistein (9),calycosin-7-O-β-D-glucopyranoside (10),genistin (11),glycitin (12),tiliroside (13).CONCLUSION Compounds 5,8,13 are isolated from genus Astragalus for the first time,and compound 2 is first isolated from this plant.

This article was aimed to study the immunomodulatory effect of ethanol sediments of the seeds of Descurainia sophia(L.) Webb. ex Prantl. both in vitro and in vivo. The lymphocyte proliferation test in vitro was carried out to explore the effect of the ethanol sediments on the proliferation of T cell and B cell in the spleen of normal mice. And, the carbon clearance test, serum hemolysin test, and delayed-type hypersensitivity test were used to investigate the influence of fraction on non-specific immunity, humoral immunity and cellular immunity in the immunosuppressive mice induced by cyclophosphamide. Besides, the immunosuppressive model was used to evaluate the effect of fraction on immune organs and content of cellular factors in blood serum. The results showed that the ethanol sediments promoted Concanavalin A (Con A) induced T cell and Lipopolysaccharides (LPS) induced B cell (P < 0.01). It increased the carbon clearance index K, phagocytic index α, half value hemolysis (HC50), and swelling degree of auricula (P < 0.05 or P < 0.01). It reduced the body weight and atrophy of thymus and spleen index (P < 0.05 or P < 0.01). It increased the contents of IL-2, IFN-γ and IL-4 in serum in immunosuppressive mice (P < 0.05 or P < 0.01). It was concluded that ethanol sediments of the seeds of D. sophia(L.) Webb. ex Prantl. can boost the lymphocyte proliferation, protect the immune organs, and enhance the non-specific and specific immunity in immunosuppressive mice, which indicated that it had immune-promotion effect.

An analytical liquid-liquid extraction-gas chromatography–mass spectrometry (LLE-GC-MS) method was established for the determination of genotoxic impurities including methyl methanesulfonate (MMS), ethyl methanesulfonate (EMS) and isopropyl methanesulfonate (IMS) in methanesulfonic acid. An Agilent HP-1MS capillary column (30 m × 0.32 m, 1 μm) was used for separating the analytes by programmed heating with the inlet temperature of 220 °C. Mass spectrometry was operated in positive ion mode, and selective ion monitors were set at m/z 80 for MMS, m/z 79 for EMS, m/z 123 for IMS and m/z 56 for internal standard butyl methanesulfonate (BMS). Results showed that the baseline separation of MMS, EMS and IMS was achieved, and the blank extraction solution had no interference; good linearity was achieved in the range of 37-1 480 ng/mL for three alkyl methanesulfonates; The mean recoveries of MMS, EMS, IMS were 104.99%, 107.26%,108.85%, respectively, with RSD ≤ 4.54%. The established method has the characteristics of specific, sensitive, accurate, stable and good versatility, and has been used for the detection and control of alkyl methanesulfonate impurities in methanesulfonic acid from a variety of manufacturers.

Objective To explore the clinical features and genetic etiology of children with cystinuria with onset of kidney stone. Methods The clinical data of 3 children with cystinuria with onset of kidney stone and the gene analysis results of SLC3A1 and SLC7A9 by PCR sequencing were retrospectively analyzed.Results Three male children were from three unrelated families, kidney stone were presented in 2 cases at 1 year old and 1 case at 14 years old. The blood amino acid spectrum was normal in all 3 cases, while the free carnitine were decreased. The urinary amino acid spectrum indicated that cystine, ornithine, arginine,and threonine increased.Gene analysis confirmed that 1 case had homozygous mutations of SLC7A9 gene c.325G>A, and his parents were carriers of c.325G>A heterozygous mutation;other 2 cases had heterozygous mutations of SLC3A1 gene, c.1365delG and c.1113C>A heterozygous mutation in one case, and c.1897_1898insTA and c.1093C>T heterozygous mutation in one case, and their parents were heterozygous mutation carriers. After treatment with potassium citrate and L-carnitine, the conditions were improved in all cases. Conclusions Inherited metabolic disease should be considered for children with kidney stone. Urine amino acid analysis and gene detection are important methods for the diagnosis of cystinuria.